α-synuclein aggregate binders and imaging methods

Abstract

Provides an α-synuclein aggregate binder with high binding selectivity to α-synuclein aggregates. An α-synuclein aggregate binder comprising: a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof (in formula (I), R1 and R2 are each independently selected from hydrogen, alkyl, alkenyl, acyl, and hydroxyalkyl; R3 is hydrogen or halogen; ring A is a benzene ring or a pyridine ring; ring B is of formula (i) or (ii); and R4 and R5 are each independently selected from hydrogen, hydroxyl, alkoxy, haloalkoxy, halohydroxyalkoxy, and aminoalkyl).

Description

Technical Field

[0001] This invention relates to α-synuclein aggregate binding agents, compositions for optical imaging of α-synuclein aggregates, compositions for radiographic imaging of α-synuclein aggregates, diagnostic agents for diseases related to α-synuclein aggregates, or companion diagnostic agents for the treatment or prevention of said diseases, diagnostic kits for diseases related to substances accumulated in the brain, optical imaging methods, radiographic imaging methods, screening methods for treatments or preventions of diseases related to α-synuclein aggregates in the brain, methods for quantifying or determining the accumulation of α-synuclein aggregates in the brain, methods for classifying and determining the accumulation of substances accumulated in the brain, α-synuclein aggregation inhibitors, and novel compounds. Background Technology

[0002] Alpha synuclein aggregates constitute the core pathology of Parkinson's disease, Lewy body dementia (DLB), and multiple system atrophy (MSA), and have a close causal relationship with neurodegeneration. Diagnosis of these diseases relies on the presence of alpha synuclein aggregates in brain anatomical pathological analysis, thus pre-mortem diagnosis is impossible. However, if alpha synuclein aggregates in the brain of an organism can be visualized, it may be possible to obtain near-confirmatory information (diagnosis) related to the diagnosis of these diseases at an early stage. Furthermore, if alpha synuclein aggregates can be visualized in the brains of disease model animals, imaging over time may aid in the evaluation of the efficacy of candidate drugs for treatment or prevention targeting alpha synuclein aggregates.

[0003] Examples of known PET (positron emission tomography) probes that bind to α-synuclein in the brain of living organisms include […]. 11 C] BF-227. But [ 11 C] BF-227 has insufficient affinity for α-synuclein aggregates, and in these diseases, only α-synuclein lesions can be detected in some MSA patients. Additionally, [ 11 C]BF-227 has the problem of nonspecific brain accumulation and low binding selectivity to α-synuclein aggregates due to its binding to amyloid β aggregates.

[0004] The inventors have developed a novel compound for imaging (visualizing) Tau protein accumulation in the brain (see Patent Document 1). The compound described in Patent Document 1 is capable of imaging Tau protein accumulation in the brain; therefore, the technology of Patent Document 1 can be applied to the treatment and prevention of diseases caused by Tau protein accumulation, such as Alzheimer's disease (AD). However, Patent Document 1 does not describe binding to α-synuclein aggregates.

[0005] (Existing technical literature)

[0006] (Patent Documents)

[0007] Patent Document 1: International Publication No. 2014 / 097474 Summary of the Invention

[0008] (The problem the invention aims to solve)

[0009] The inventors have discovered that in the compound described in Patent Document 1, (2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol (PBB3) can bind to α-synuclein aggregates with a certain affinity.

[0010] However, the binding selectivity of PBB3 to α-synuclein aggregates is lower than that of PBB3 to Tau aggregates. Therefore, PBB3 is not very suitable as an α-synuclein aggregate probe. Thus, a α-synuclein aggregate binder with high binding selectivity is desired.

[0011] Furthermore, if it were possible not only to detect and diagnose α-synuclein aggregates, but also to inhibit (block) their aggregation, then diseases caused by α-synuclein aggregates could be prevented or treated. Therefore, α-synuclein aggregation inhibitors that can inhibit α-synuclein aggregation are anticipated.

[0012] In view of the above, the present invention was made. The object of the present invention is to provide an α-synuclein aggregate binder with high binding selectivity to α-synuclein aggregates, an imaging method using the α-synuclein aggregate binder, a new compound that can be used with the α-synuclein aggregate binder, and an α-synuclein aggregation inhibitor.

[0013] (Technical means used to solve the problem)

[0014] The inventors discovered that among the compounds described in Patent Document 1, the compound of formula (I) having a butenyl structure and a benzothiazole structure or a benzofuran structure exhibits high binding selectivity to α-synuclein aggregates. Through further repeated research, this invention was completed. More specifically, the present invention provides the following solution.

[0015] [1] A binder for α-synuclein aggregates, comprising:

[0016] The compound represented by formula (I), its pharmaceutically permissible salt, or its solvate,

[0017]

[0018] (In formula (I),

[0019] R1 and R2 are each independently selected from hydrogen, alkyl, alkenyl, acyl, and hydroxyalkyl.

[0020] R3 is hydrogen or halogen.

[0021] Ring A is a benzene ring or a pyridine ring.

[0022] Ring B is represented by either equation (i) or equation (ii).

[0023]

[0024] R4 and R5 are each independently selected from hydrogen, hydroxyl, alkoxy, haloalkoxy, halohydroxyalkoxy, and aminoalkyl.

[0025] [2] According to the binder described in [1], wherein ring A is a pyridine ring.

[0026] [3] According to the binder described in [1] or [2], wherein ring B is of formula (i).

[0027] [4] According to the binder of [1], wherein the compound represented by formula (I) is selected from the group consisting of:

[0028]

[0029]

[0030] [5] The binder according to any one of [1] to [4], wherein,

[0031] One or more atoms of the compound represented by formula (I) are radioactive isotopes of that atom.

[0032] [6] A composition for optical imaging of α-synuclein aggregates, comprising:

[0033] The binder as described in any one of [1] to [4].

[0034] [7] A composition for radiographic imaging of α-synuclein aggregates, comprising:

[0035] [5] The binder described above.

[0036] [8] A diagnostic drug comprising any one of [1] to [5],

[0037] This diagnostic drug is a diagnostic drug for diseases associated with α-synuclein aggregates, or a companion diagnostic drug for the treatment or prevention of said diseases.

[0038] [9] A diagnostic kit for a disease related to substances accumulating in the brain, comprising:

[0039] The binder selected from any one of [1] to [5]; and the binder selected from 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-ol, 2-((1E,3E)-4-(6-((11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-ol, 1-fluoro-3-( At least one of 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol and 1-(18)F-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol).

[0040]

[10] An optical imaging method for α-synuclein aggregates in the brain, comprising the following steps:

[0041] The brain of a test organism, which has been given any one of the binders in [1] to [4], is irradiated with light of a first wavelength from outside the brain, and then light of a second wavelength, different from the first wavelength, emitted from the brain is detected.

[0042]

[11] A radiographic imaging method for α-synuclein aggregates in the brain, comprising the following steps:

[0043] Detect radiation emitted from the brain of a test organism that has been given the binding agent described in [5].

[0044]

[12] A method for screening drugs for the treatment or prevention of diseases associated with α-synuclein aggregates in the brain, comprising the following steps:

[0045] The candidate substance is screened based on the difference in the amount and / or distribution of light or radiation detected in the test organism before and after the administration of the candidate substance by the imaging method described in

[10] or

[11] .

[0046]

[13] A method for quantifying or determining the accumulation of α-synuclein aggregates in the brain.

[0047] It includes the following steps: detecting radiation emitted from the brain of a test organism to which the binder described in [5] has been administered, and,

[0048] Based on the amount and / or distribution of the detected radiation, the accumulation of α-synuclein aggregates in the brain can be quantified or determined.

[0049]

[14] A method for classifying and determining the accumulation of substances in the brain, comprising:

[0050] Step 1: Detect radiation emitted from the brain of a test organism that has been given the binding agent described in [5];

[0051] Step 2 involves detecting radiation emitted from the brain of the test organism that was exposed at a different time than in Step 1 to at least one of 2-((1E,3E)-4-(6-((11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol and 1-(18)F-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-yloxy)propane-2-ol, and,

[0052] Based on the radiation quantity and / or distribution data detected in step 1 and the radiation quantity and / or distribution data detected in step 2, the classification and accumulation of substances accumulated in the brain are determined.

[0053]

[15] An α-synuclein aggregation inhibitor, comprising:

[0054] The compound represented by formula (I), its pharmaceutically permissible salt, or its solvate,

[0055]

[0056] (In formula (I),

[0057] R1 and R2 are each independently selected from hydrogen, alkyl, alkenyl, acyl, and hydroxyalkyl.

[0058] R3 is hydrogen or halogen.

[0059] Ring A is a benzene ring or a pyridine ring.

[0060] Ring B is represented by either equation (i) or equation (ii).

[0061]

[0062] R4 and R5 are each independently selected from hydrogen, hydroxyl, alkoxy, haloalkoxy, halohydroxyalkoxy, and aminoalkyl.

[0063]

[16] An α-synuclein aggregation inhibitor as described in

[15] , wherein ring A is a pyridine ring.

[0064]

[17] An α-synuclein aggregation inhibitor according to

[15] or

[16] , wherein ring B is of formula (i).

[0065]

[18] According to the α-synuclein aggregation inhibitor described in

[15] , wherein,

[0066] The compounds represented by formula (I) are selected from the following group:

[0067]

[0068]

[0069]

[19] A compound represented by the following formula, a pharmaceutically permissible salt thereof, or a solvate thereof,

[0070]

[20] A compound represented by the following formula, a pharmaceutically permissible salt thereof, or a solvate thereof,

[0071] (In the formula, at least one of the atoms marked with * is a radioactive isotope of that atom.)

[0072]

[21] An intermediate shown in the following formula, used in the synthesis of the compound shown in

[19] ,

[0073]

[0074]

[22] An intermediate shown in the following formula, used in the synthesis of the compound shown in

[20] ,

[0075]

[0076]

[0077] (The effect of the invention)

[0078] According to the present invention, an α-synuclein aggregate binder with high binding selectivity to α-synuclein aggregates can be provided.

[0079] Furthermore, it is possible to provide an imaging method that uses this α-synuclein aggregate binder.

[0080] In addition, it can provide new compounds that can be used as α-synuclein aggregate binders and for other applications.

[0081] In addition, it can inhibit the aggregation of α-synuclein. Attached Figure Description

[0082] Figure 1A The results of fluorescence microscopy measurements of the brains of DLB ​​patients and AD patients are shown.

[0083] Figure 1BThe results of fluorescence microscopy measurements of the brains of DLB ​​patients and AD patients are shown.

[0084] Figure 2 The quantitative results show the fluorescence intensity of the lesion area and the non-lesion area.

[0085] Figure 3 The results of fluorescence microscopy measurements in mice after inoculation with α-synuclein fibers are shown.

[0086] Figure 4 The results of two-photon laser scanning fluorescence microscopy examination of model mice 6 weeks after injection of α-synuclein fibrillary solution are shown.

[0087] Figure 5 The PET imaging results are shown.

[0088] Figure 6 The PET imaging results are shown.

[0089] Figure 7 The results of fluorescence microscopy examination of mice after inoculation with α-synuclein fibers are shown.

[0090] Figure 8 The PET imaging results are shown.

[0091] Figure 9 The PET imaging results are shown.

[0092] Figure 10 The PET imaging results are shown.

[0093] Figure 11 The PET imaging results are shown.

[0094] Figure 12 The PET imaging results are shown.

[0095] Figure 13 This demonstrates the inhibitory effect of α-synuclein aggregation.

[0096] Figure 14 This demonstrates the inhibitory effect of α-synuclein aggregation. Detailed Implementation

[0097] <Definition>

[0098] The term "alkyl" refers to a monovalent group formed by the loss of one hydrogen atom from an aliphatic saturated hydrocarbon. Alkyl groups have, for example, 1 to 15 carbon atoms, typically 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 2 to 6 carbon atoms. Alkyl groups can be straight-chain, branched, or cyclic. Alkyl groups are not limited to any particular group; examples include methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, and hexyl. Alkyl groups can also be substituted with suitable substituents.

[0099] In this specification, the number of carbon atoms (1-15, 1-10, 1-8, 1-6, 1-5, 1-4, 1-3, 1-2, 2-8, 2-6, 2-4, 3-8, 3-6, 4-8, 4-6) are respectively denoted as C. 1-15 C 1-10 C 1-8 C 1-6 C 1-5 C 1-4 C 1-3 C 1-2 C 2-8 C 2-6 C 2-4 C 3-8 C 3-6 C 4-8 and C 4-6 wait.

[0100] The term "cycloalkyl" refers to a monovalent group formed by the loss of one hydrogen atom from an aliphatic saturated hydrocarbon that forms a carbocyclic ring. Cycloalkyl groups typically have 3 to 10 carbon atoms, and more commonly 3 to 8, 3 to 6, 3 to 5, 3 to 4, 4 to 5, 4 to 6, or 4 to 8 carbon atoms. The type of cycloalkyl group is not limited; examples include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, and cyclooctane. Cycloalkyl groups can also be substituted with suitable substituents.

[0101] The term "alkenyl" refers to an aliphatic unsaturated hydrocarbon group having at least one double bond. Alkenyl groups have, for example, 2 to 15 carbon atoms, typically 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 6, 3 to 8, 4 to 6, 4 to 7, or 4 to 8 carbon atoms. Alkenyl groups can be straight-chain or branched. The alkenyl group is not limited; specific examples include vinyl (i.e., -CH=CH2), allyl (i.e., -CH2CH=CH2), -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, 1,3-butadienyl (i.e., -CH=CH-CH=CH2), and hepta-1,6-dien-4-yl (i.e., -CH2-(CH2CH=CH2)2), etc. The alkenyl group can also be substituted with suitable substituents.

[0102] The term "alkynyl" refers to an aliphatic unsaturated hydrocarbon group having at least one triple bond. Alynyl groups typically have 2 to 15 carbon atoms, including 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 6, 4 to 6, 4 to 7, or 4 to 8 carbon atoms. Alynyl groups can be straight-chain or branched. There is no limitation on the type of alkynyl group; examples include ethynyl (i.e., -C≡CH), -C≡CH(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and -CH2C≡C(CH2CH3). Alynyl groups can also be substituted with suitable substituents.

[0103] The term "acyl" refers to a group represented by -CO-R. Here, R is, for example, an alkyl, alkenyl, or alkynyl group. Acyl groups are not limited to any particular group and can include, for example, acetyl (i.e., -COCH3), ethyl carbonyl, propyl carbonyl, pentyl carbonyl, cyclohexyl carbonyl, octyl carbonyl, 2-ethylhexyl carbonyl, dodecyl carbonyl, phenyl carbonyl, benzyl carbonyl, naphthyl carbonyl, and pyridyl carbonyl. Acyl groups can also be substituted with suitable substituents.

[0104] The term "hydroxyl" or "hydroxyl group" refers to -OH.

[0105] The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group (i.e., -OH). Hydroxyalkyl groups are not limited to a single type; examples include hydroxymethyl (i.e., -CH2OH), 2-hydroxyethyl (i.e., -CH2CH2OH), 1-hydroxyethyl (i.e., -CH(OH)CH3), 3-hydroxypropyl (i.e., -CH2CH2CH2OH), 2-hydroxypropyl (i.e., -CH2CH(OH)CH3), and 1-hydroxypropyl (i.e., -CH(OH)CH2CH3). Hydroxyalkyl groups can also be substituted with suitable substituents.

[0106] The term "halogen" or "halogen" refers to fluorine (-F), chlorine (-Cl), bromine (-Br), and iodine (-I).

[0107] The term "alkoxy" refers to an alkyl group (i.e., -O-alkyl) bonded to other groups separated by an oxygen atom. Alkoxy groups are not limited to any particular group; examples include methoxy (i.e., -O-methyl), ethoxy (i.e., -O-ethyl), propoxy (i.e., -O-propyl), -O-isopropyl, -O-2-methyl-1-propyl, -O-2-methyl-2-propyl, -O-2-methyl-1-butyl, -O-3-methyl-1-butyl, -O-2-methyl-3-butyl, -O-2,2-dimethyl-1-propyl, -O-2-methyl-1-pentyl, 3... -O-methyl-1-pentyl, -O-4-methyl-1-pentyl, -O-2-methyl-2-pentyl, -O-3-methyl-2-pentyl, -O-4-methyl-2-pentyl, -O-2,2-dimethyl-1-butyl, -O-3,3-dimethyl-1-butyl, -O-2-ethyl-1-butyl, -O-butyl, -O-isobutyl, -O-tert-butyl, -O-pentyl, -O-isopentyl, -O-neopentyl, and -O-hexyl. The alkoxy group can also be substituted with suitable substituents.

[0108] The term "haloalkyl" refers to an alkyl group that has been substituted with at least one halogen. Examples of haloalkyl groups include fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl. The list of haloalkyl groups is not limited and includes fluoromethyl, chloromethyl, bromomethyl, iodomethyl, fluoroethyl, chloroethyl, bromoethyl, iodoethyl, fluoropropyl, chloropropyl, bromopropyl, iodopropyl, fluorobutyl, chlorobutyl, bromobutyl, iodobutyl, fluoropentyl, chloropentyl, bromopentyl, iodopentyl, fluorohexyl, chlorohexyl, bromohexyl, fluoroheptyl, chloroheptyl, bromoheptyl, fluorooctyl, chlorooctyl, bromooctyl, and iodooctyl, etc. Haloalkyl groups can also be substituted with suitable substituents.

[0109] The term "haloalkoxy" refers to an alkoxy group that has been substituted with at least one halogen (i.e., -O-haloalkyl). Haloalkoxy groups include fluoroalkoxy, chloroalkoxy, bromoalkoxy, and iodoalkoxy.

[0110] The term "halohydroxyalkoxy" refers to a haloalkoxy group that has been substituted with a hydroxyl group. Examples of halohydroxyalkoxy groups include fluorohydroxyalkoxy, chlorohydroxyalkoxy, bromohydroxyalkoxy, and iodohydroxyalkoxy. Examples of halohydroxyalkoxy groups include -O-CH(F)(OH), -O-CH2CH(F)(OH), -O-CH(OH)-CH2(F), -O-CH2-CH(F)(OH), -O-CH(OH)-CH2-CH2(F), -O-CH2-CH(OH)-CH2(F), -O-CH(CH2-F)(CH2OH), and -O-CH2-CH2-CH(F)(OH).

[0111] The term "nitro" refers to -NO2.

[0112] The term "amino" refers to -NH2.

[0113] The term "aminoalkyl" refers to an alkyl group that has been substituted with an amino group. There is no limitation on the type of aminoalkyl group; examples include aminomethyl, aminoethyl, aminopropyl, aminoisopropyl, aminobutyl, aminopentyl, aminohexyl, and aminooctyl.

[0114] The term "substituent" refers to one or more atoms or groups of atoms introduced into a chemical structural formula. Examples of substituents include C. 1-8 Alkyl groups (methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, or n-hexyl or their isomers, etc.), C 2-8 Alkenyl (vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), and -C(CH2CH3)=CH2, etc.), C 2-8 Alkyne (ethynyl, -C≡CH(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and -CH2C≡C(CH2CH3), etc.), alkoxy, hydroxyl, halogen, haloalkyl, cycloalkyl (cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, etc.), amino, nitro, acyl (acetyl (i.e., -COCH3), etc.), carboxyl (i.e., -COOH), ester (i.e., -COOR) x , where R x It is C 1-6 Alkyl groups, etc.), amide groups (i.e., -CONR) y R z Among them, R y and R z Independently H or C 1-6Alkyl groups, mercapto groups (i.e., -SH), sulfonic acid groups (i.e., -SO3H), nitrile groups (i.e., -CN), aromatic rings (aryl, phenyl, benzyl, or naphthyl, etc.), heterocyclic groups (pyrrolidinyl, tetrahydrofuranyl, pyrrolidinyl, furanyl, thiophenyl, piperidinyl, aniline carbonyl (oxanyl, or pyridinyl, etc.), etc.

[0115] The term "medically permissible salt" refers to a salt that is harmless to mammals, especially humans. Pharmaceutically permissible salts can be formed using non-toxic acids or bases containing inorganic acids or bases, or organic acids or bases. Examples of pharmaceutically permissible salts include: metal salts formed from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc; or organic salts formed from lysine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (i.e., N-methylglucosamine), and procaine. Furthermore, pharmaceutically permissible salts include acid addition salts and base addition salts.

[0116] The term "pharmaceutical-permitted carrier" refers to physiological saline solutions; liquid or solid fillers, diluents, solvents, or encapsulating materials, etc., that are pharmaceutically permissible materials, compositions, or excipients. Examples of pharmaceutically permissible carriers include water, saline, physiological saline or phosphate-buffered saline (PBS), sodium chloride injection, Ringer's solution, isotonic glucose injection, sterile water injection, glucose, and lactated Ringer's solution.

[0117] The term "effective amount" refers to the amount of a compound or composition that is sufficient to achieve the desired effect. For example, in some embodiments, an effective amount refers to the amount of a compound or composition that enables successful optical and radiographic imaging of brain accumulations such as α-synuclein aggregates.

[0118] The term "solvent" refers to a solvent-containing compound formed by the association of one or more solvent molecules with a compound. Solvents include, for example, monosolvents, disolvents, trisolvents, and tetrasolvents. Additionally, solvates include hydrates.

[0119] The term "hydrate" refers to a compound or its salt that further comprises stoichiometric or non-stoichiometric amounts of water bound by non-covalent intermolecular forces. Hydrates include, for example, monohydrates, dihydrates, trihydrates, and tetrahydrates.

[0120] The term "treatment" refers to reducing or improving the severity, degree, and / or duration of a disease or condition.

[0121] The term “prevention” refers to reducing the risk of developing or worsening a given disease or condition, or reducing or suppressing the recurrence, onset, or worsening of symptoms of a given disease or one or more conditions.

[0122] <α-synuclein aggregate binder>

[0123] The α-synuclein aggregate binder of the present invention (hereinafter also referred to as binder) comprises: a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof.

[0124] The compound represented by formula (I) is based on the compound described in Patent Document 1, but instead of having a butene structure, it has a butene structure and either a benzothiazole structure or a benzofuran structure. It is known that the compound represented by formula (I) has a higher binding selectivity relative to α-synuclein aggregates compared to the binding selectivity relative to Tau protein and amyloid β aggregates. Furthermore, similarly, the compound represented by formula (I) also exhibits higher binding selectivity relative to α-synuclein aggregates in pharmaceutically permissible salts and in solvates of the compound represented by formula (I) compared to the binding selectivity relative to Tau protein and amyloid β aggregates.

[0125] Furthermore, the compound shown in formula (I) emits fluorescence. Additionally, one or more atoms in the compound shown in formula (I) may be radioactive isotopes of those atoms.

[0126] Therefore, the binder of the present invention can be used as a molecular probe for optical and radiographic imaging of α-synuclein aggregates accumulated in the brain.

[0127] In addition, as described above, the compound represented by formula (I), its pharmaceutically permissible salt, and its solvates have a high binding selectivity to α-synuclein aggregates, and thus can inhibit the aggregation of α-synuclein, as will be described in detail later.

[0128] Alpha synuclein is also present in some neuronal synapses in the normal brain. Alpha synuclein aggregates are formed by the aggregation of alpha synuclein.

[0129]

[0130] (In formula (I),

[0131] R1 and R2 are each independently selected from hydrogen, alkyl, alkenyl, acyl, and hydroxyalkyl.

[0132] R3 is hydrogen or halogen.

[0133] Ring A is a benzene ring or a pyridine ring.

[0134] Ring B is represented by either equation (i) or equation (ii).

[0135]

[0136] R4 and R5 are each independently selected from hydrogen, hydroxyl, alkoxy, haloalkoxy, halohydroxyalkoxy, and aminoalkyl.

[0137] In one embodiment, ring B is equation (i). In other embodiments, ring B is equation (ii).

[0138] If ring B is of formula (i), then R4 and R5 can be present at substituted positions on the benzothiazole ring of formula (i). Preferably, R4 and R5 are present at positions 6 and 5 of the benzothiazole ring of formula (i), respectively. If ring B is of formula (ii), then R4 and R5 can be present at substituted positions on the benzofuran ring of formula (ii). Preferably, R4 and R5 are present at positions 5 and 6 of the benzofuran ring of formula (ii), respectively.

[0139] In one embodiment, ring A is a pyridine ring. In other embodiments, ring A is a benzene ring. Preferably, in the direction of the structural formula represented by formula (I), ring A is a pyridine ring as shown in the following structural formula.

[0140]

[0141] In one embodiment, both R1 and R2 are hydrogen.

[0142] In one embodiment, R1 and R2 are each independently hydrogen or alkyl, especially C 1-8 Alkyl, preferably methyl. In other embodiments, R1 is hydrogen and R2 is an alkyl group, especially C. 1-6 Alkyl groups, preferably methyl groups. In other embodiments, both R1 and R2 are alkyl groups, preferably both C1 and C2. 1-6 Alkyl groups, more preferably methyl groups.

[0143] In one embodiment, R1 and R2 are each independently hydrogen or alkenyl, especially C. 2-8 The alkenyl group is preferably allyl (i.e., -CH2CH=CH2) or hept-1,6-dien-4-yl (i.e., -CH2-(CH2CH=CH2)2). In other embodiments, R1 is hydrogen and R2 is an alkenyl group, especially C. 2-8 The alkenyl group is preferably allyl (i.e., -CH2CH=CH2) or hept-1,6-dien-4-yl (i.e., -CH2-(CH2CH=CH2)2). In other embodiments, R1 and R2 are both alkenyl groups, preferably both C. 2-8 Alkenyl groups, more preferably allyl (i.e., -CH2CH2CH=CH2) or hept-1,6-dien-4-yl (i.e., -CH2-(CH2CH=CH2)2).

[0144] In one embodiment, R1 and R2 are each independently hydrogen or acyl, especially C. 1-8The acyl group, preferably an acetyl group (i.e., -COCH3). In other embodiments, R1 is hydrogen and R2 is an acyl group, especially C. 1-8 The acyl group is preferably an acetyl group (i.e., -COCH3). In other embodiments, both R1 and R2 are acyl groups, preferably both C. 1-8 Acyl groups, more preferably acetyl groups (i.e., -COCH3).

[0145] In one embodiment, R1 and R2 are each independently hydrogen or hydroxyalkyl, especially hydroxyl C. 1-8 Alkyl group, preferably hydroxypropyl, more preferably 3-hydroxypropyl (i.e., -CH2CH2CH2OH). In other embodiments, R1 is hydrogen and R2 is hydroxyalkyl, especially hydroxyC 1-8 Alkyl group, preferably hydroxypropyl, more preferably 3-hydroxypropyl (i.e., -CH2CH2CH2OH). In other embodiments, R1 and R2 are both hydroxyalkyl groups, preferably both hydroxyC. 1-8 Alkyl groups, more preferably hydroxypropyl, and even more preferably 3-hydroxypropyl (i.e., -CH2CH2CH2OH).

[0146] In one embodiment, R3 is hydrogen. In other embodiments, R3 is a halogen, i.e., F, Cl, Br, or I. Preferably, R3 is F. More preferably, R3 is... 18 F. Preferably, in the direction of the structural formula represented by equation (I), R3 exists in the following positions.

[0147]

[0148] Preferably, in the direction of the structural formula represented by equation (I), the relationship between ring A and R3 is as follows.

[0149]

[0150] In one embodiment, both R4 and R5 are hydrogen.

[0151] In one embodiment, R4 and R5 are each independently hydrogen or hydroxyl. In other embodiments, R4 is hydroxyl and R5 is hydrogen. In yet another embodiment, R4 is hydrogen and R5 is hydroxyl. In still another embodiment, both R4 and R5 are hydroxyl.

[0152] In one embodiment, R4 and R5 are each independently hydrogen or alkoxy, especially methoxy. In other embodiments, R4 is alkoxy, especially methoxy, and R5 is hydrogen. In still other embodiments, R4 is hydrogen and R5 is alkoxy, especially methoxy. In yet another embodiment, both R4 and R5 are alkoxy, preferably both are methoxy.

[0153] In one embodiment, R4 and R5 are each independently hydrogen or a haloalkoxy group, especially fluoroalkoxy, chloroalkoxy, bromoalkoxy, or iodoalkoxy. In other embodiments, R4 is a haloalkoxy group, especially fluoroalkoxy, chloroalkoxy, bromoalkoxy, or iodoalkoxy, and R5 is hydrogen. In further embodiments, R4 is hydrogen, and R5 is a haloalkoxy group, especially fluoroalkoxy, chloroalkoxy, bromoalkoxy, or iodoalkoxy. In still other embodiments, both R4 and R5 are haloalkoxy groups, especially fluoroalkoxy, chloroalkoxy, bromoalkoxy, or iodoalkoxy. In one embodiment, the fluoroalkoxy group contains a radioactive isotope.

[0154] In one embodiment, R4 and R5 are each independently hydrogen or halohydroxyalkoxy, especially fluorohydroxyalkoxy, preferably fluorohydroxyC. 1-3 The alkoxy group is more preferably -O-CH2-CH(OH)-CH2(F) or -O-CH(CH2-F)(CH2OH). In other embodiments, R4 is a halohydroxyalkoxy group, especially a fluorohydroxyalkoxy group, preferably a fluorohydroxyC group. 1-3 The alkoxy group is more preferably -O-CH2-CH(OH)-CH2(F) or -O-CH(CH2-F)(CH2OH), and R5 is hydrogen. In other embodiments, R4 is hydrogen, and R5 is a halohydroxyalkoxy group, especially a fluorohydroxyalkoxy group, preferably a fluorohydroxyalkoxy group. 1-3 The alkoxy group is more preferably -O-CH2-CH(OH)-CH2(F) or -O-CH(CH2-F)(CH2OH). In other embodiments, R4 and R5 are both halohydroxyalkoxy groups, more preferably fluorohydroxyalkoxy groups, and even more preferably fluorohydroxyC groups. 1-3 The alkoxy group, and more preferably -O-CH2-CH(OH)-CH2(F) or -O-CH(CH2-F)(CH2OH), is present. In one embodiment, the fluorohydroxyalkoxy group contains a radioactive isotope. Preferably, the fluorohydroxyalkoxy group is -O-CH2-CH(OH)-CH2(F). 18 F), or -O-CH(CH2- 18 F)(CH2OH).

[0155] In one embodiment, R4 and R5 are each independently hydrogen or an aminoalkyl group, especially aminomethyl or aminoethyl. In other embodiments, R4 is an aminoalkyl group, especially aminomethyl or aminoethyl, and R5 is hydrogen. In other embodiments, R4 is hydrogen and R5 is an aminoalkyl group, especially aminomethyl or aminoethyl. In other embodiments, both R4 and R5 are aminoalkyl groups, preferably both aminomethyl or aminoethyl.

[0156] In one embodiment, the compound represented by formula (I) is the compound represented by formula (II).

[0157]

[0158] (In the formula, R1, R2, R3, R4, and R5 are the same as R1, R2, R3, R4, and R5 in formula (I), respectively.)

[0159] In one embodiment, the compound represented by formula (I) is the compound represented by formula (III).

[0160]

[0161] (In the formula, R1, R2, R3, R4, and R5 are the same as R1, R2, R3, R4, and R5 in formula (I), respectively.)

[0162] In one embodiment, the compound represented by formula (I) is the compound represented by formula (IV).

[0163]

[0164] (In the formula, R1, R2, R3, R4, and R5 are the same as R1, R2, R3, R4, and R5 in formula (I), respectively.)

[0165] In one embodiment, the compound represented by formula (I) is the compound represented by formula (V).

[0166]

[0167] (In the formula, R1, R2, R3, R4, and R5 are the same as R1, R2, R3, R4, and R5 in formula (I), respectively.)

[0168] In one embodiment, in the compounds of formulas (I) to (V), one or more atoms are radioactive isotopes of that atom. The radioactive isotopes are selected from... 15 O、 13 N、 11 C, and 18 F, etc., are not particularly limited. Preferably, the radioactive isotope is... 11 C or 19 F. Among these, considering 11 The half-life of C is approximately 20 minutes. 18 The half-life of F is approximately 110 minutes, therefore, by 18 F-labeled compounds have higher commercial value. Therefore, most preferably, the radioactive isotope is... 18 F.

[0169] Preferably, any one or more of R1 to R5 is a base containing a radioactive isotope. More preferably, R1 and / or R2 is a base containing a radioactive isotope, for example, a base containing... 11 The base of C (containing 11 CH3 of [ 11 (C) Alkyl group, etc. More preferably, R3 is a group containing a radioactive isotope, for example, - 18 F. More preferably, R4 and / or R5 are bases containing radioactive isotopes, for example, bases containing... 11 The base of C (containing -O) 11 CH3 of [ 11 C] alkoxy group, etc.), or containing 18 The F group (-O-CH2-CH(OH)-CH2( 18 F), and containing -O-CH(CH2- 18 F)(CH2OH) of [ 18 F] Fluorohydroxyalkoxy, etc.). Among them, [ 11 [C] An alkyl group refers to an alkyl group in which one or more carbon atoms are... 11 C. [ 11 C] An alkoxy group refers to an alkoxy group consisting of one or more carbon atoms that are... 11 C. [ 18 F] Fluorohydroxyalkoxy refers to 18 The group formed by F bonding with hydroxyalkoxy groups.

[0170] As specific examples of compounds represented by formula (I), the following compounds can be cited.

[0171]

[0172]

[0173]

[0174]

[0175] In one embodiment, in the specific compound described above, one or more atoms are radioactive isotopes of that atom. Preferably, in the aminobenzyl or aminopyridyl group bonded to the benzene or pyridine ring, the carbon atom bonded to the nitrogen atom of the amino group is a radioactive isotope. 11 C. Preferably, F in the above-mentioned specific compound is a radioactive isotope. 18 F. Preferably, the carbon atom of the methoxy group bonded to the benzothiazole ring is a radioactive isotope. 11C. Furthermore, preferably, the atom marked with * in the specific compound structural formula above (if there are two * marks in the structural formula, it refers to any one or both of them) is a radioactive isotope of that atom, for example... 11 C or 18 F. In this specification, [ 11 The meaning of names such as C05-04, etc., is that the atoms marked with an asterisk (*) in the structural formulas numbered C05-04, etc., are... 11 C.

[0176] The method of synthesizing the compound shown in formula (I) is not particularly limited, and it can be manufactured by referring to the synthesis method described in Patent Document 1.

[0177] Furthermore, among the compounds shown in formula (I) above, the compound shown in the following formula, its pharmaceutically permissible salt, or its solvate are novel compounds. The compound shown in the following formula can be used as the aforementioned α-synuclein aggregate binding agent, or for uses other than as an α-synuclein aggregate binding agent.

[0178]

[0179]

[0180] (In the formula, at least one of the atoms marked with * is a radioactive isotope of that atom.)

[0181] For example, the above-mentioned new compounds can be synthesized using the following intermediates and the synthesis methods described in the following examples.

[0182]

[0183]

[0184] Alpha synuclein aggregate binders can be contained in pharmaceutically permissible carriers. Examples of such pharmaceutically permissible carriers include water, saline, physiological saline or phosphate-buffered saline (PBS), sodium chloride injection, Ringer's solution, isotonic glucose injection, sterile water injection, glucose, and lactated Ringer's solution.

[0185] The amount of the compound of formula (I), its pharmaceutically permissible salt, or its solvate, and the pharmaceutically permissible carrier contained in the α-synuclein aggregate binder is not particularly limited, and their amounts can be determined based on various factors such as: the type of compound used; the age, weight, health status, sex, and diet of the mammal to which the medication is administered; the frequency and route of administration; the duration of treatment; and other concurrently used drugs. The pharmaceutically permissible carrier may be present in amounts ranging from 1% to 99% by weight of the α-synuclein aggregate binder. The α-synuclein aggregate binder can be prepared such that the compound shown in formula (I) is administered at a dosage of 5 ng / kg to 5 mg / kg per unit body weight (kg) of the subject, wherein the lower limit is preferably 5 ng / kg, 0.01 mg / kg, 0.05 mg / kg and 0.1 mg / kg, and the upper limit is preferably 5 mg / kg, 3 mg / kg, 1 mg / kg and 20 μg / kg.

[0186] <A composition for optical imaging of α-synuclein aggregates>

[0187] The optical imaging composition for α-synuclein aggregates of the present invention (hereinafter also referred to as optical imaging composition) comprises the binding agent of the present invention described above. This optical imaging includes in vitro imaging, in vitro imaging, and in vivo imaging.

[0188] Examples of optical imaging methods include fluorescence microscopy, multiphoton imaging, two-photon imaging, and near-infrared fluorescence imaging.

[0189] Compositions for optical imaging may contain pharmaceutically permissible carriers. Examples of pharmaceutically permissible carriers include water, saline, physiological saline or phosphate-buffered saline (PBS), sodium chloride injection, Ringer's solution, isotonic glucose injection, sterile water injection, glucose, and lactated Ringer's solution.

[0190] The amounts of the compound of formula (I), its pharmaceutically permissible salts, or solvates thereof, and pharmaceutically permissible carriers contained in the optical imaging composition are not particularly limited, and their amounts can be determined based on various factors such as: the type of compound used; the age, weight, health status, sex, and diet of the mammal to which it is administered; the frequency and route of administration; the duration of treatment; and other concurrently used pharmaceutical agents. The amount of pharmaceutically permissible carriers can be 1 to 99% by weight of the optical imaging composition. The optical imaging composition can be prepared, for example, to administer the compound of formula (I) at an amount (mg) relative to the body weight (kg) of the subject, of 0.01 mg / kg to 5 mg / kg, preferably 0.05 mg / kg to 3 mg / kg, and more preferably 0.1 mg / kg to 1 mg / kg.

[0191] <A composition for radiographic imaging of α-synuclein aggregates>

[0192] The radiographic imaging composition for α-synuclein aggregates of the present invention (hereinafter also referred to as radiographic imaging composition) comprises the binding agent of the present invention described above. This radiographic imaging includes in vitro imaging, in vitro imaging, and in vivo imaging.

[0193] Examples of radiographic imaging techniques include, for instance, positron emission tomography (PET), single photon emission computed tomography (SPECT), and autoradiography.

[0194] Compositions for radiographic imaging may contain pharmaceutically permissible carriers. Examples of pharmaceutically permissible carriers include water, saline, physiological saline or phosphate-buffered saline (PBS), sodium chloride injection, Ringer's solution, isotonic glucose injection, sterile water injection, glucose, and lactated Ringer's solution.

[0195] The amounts of the compound of formula (I), its pharmaceutically permissible salts, or solvates thereof, and pharmaceutically permissible carriers contained in the radiographic imaging composition are not particularly limited, and their amounts can be determined based on various factors such as: the type of compound used; the age, weight, health status, sex, and diet of the mammal to which it is administered; the number of administrations and the route of administration; the duration of treatment; and other concurrently used pharmaceutical agents. The amount of pharmaceutically permissible carriers may be 1 to 99% by weight of the radiographic imaging composition. The radiographic imaging composition can, for example, be prepared such that the compound of formula (I) can be administered at an amount of 5 ng / kg to 5 mg / kg, preferably 5 ng to 20 μg / kg, relative to the body weight (kg) of the subject.

[0196] <Diagnostic drugs for diseases associated with α-synuclein aggregates, or companion diagnostic drugs for the treatment or prevention of said diseases>

[0197] The diagnostic agents for diseases related to α-synuclein aggregates of the present invention, or companion diagnostic agents for the treatment or prevention of said diseases (hereinafter also referred to as companion diagnostic agents), comprise the binding agent of the present invention described above. A companion diagnostic agent for treatment refers to a diagnostic agent used to determine whether treatment is likely to be implemented after the disease has been identified. Furthermore, a companion diagnostic agent for prevention refers to a diagnostic agent used to predict future onset of the disease or to determine whether preventive onset suppression is likely to be implemented after the prodromal symptoms of the disease have been identified.

[0198] By comparing the data on the amount and / or distribution of α-synuclein aggregates in the brain obtained from the subject using the aforementioned diagnostic drugs with the previously known correlation between the aforementioned diseases and the amount and / or distribution of α-synuclein aggregates, it is possible to make a relevant diagnosis of the aforementioned diseases in the subject (specifically, whether the subject has the aforementioned diseases, the severity, the likelihood of onset, etc.).

[0199] Furthermore, by comparing the data on the amount and / or distribution of α-synuclein aggregates in the brain obtained from the subject using the aforementioned companion diagnostic drugs with the previously known correlation between the aforementioned diseases and the amount and / or distribution of α-synuclein aggregates in the brain, it is possible to grasp the subject's disease status. Therefore, it is possible to formulate a prevention / treatment plan for the aforementioned diseases (types and combinations of prophylactic / therapeutic drugs, dosages, and methods of administration, etc.).

[0200] One embodiment of the present invention is a medicine for the treatment or prevention of diseases associated with α-synuclein aggregates, and also relates to a medicine administered according to a dosing plan based on relevant data on the amount and / or distribution of α-synuclein aggregates in the brain obtained in the aforementioned companion diagnostics.

[0201] <Diagnostic kits for diseases related to substances accumulating in the brain>

[0202] The diagnostic kit for diseases related to substances accumulating in the brain (hereinafter also referred to as the diagnostic kit) of the present invention comprises: the binding agent of the present invention described above; and a mixture selected from 2-((1E,3E)-4-(6-(11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol, 2-((1E,3E)-4-(6-((11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol, 2- ...11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol, 2-(1E,3E)-4-(6-(11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol, 2-(1E,3E)-4-(6-(11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol, 2- At least one of d]thiazol-6-ol, 1-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol and 1-(18)F-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol.

[0203] Examples of substances that accumulate in the brain include α-synuclein aggregates, Tau protein, and amyloid-β aggregates.

[0204] Diseases associated with substances accumulating in the brain include: diseases associated with α-synuclein aggregates, such as Parkinson's disease, Lewy body dementia (DLB), and multiple system atrophy (MSA); and diseases associated with aggregates of Tau protein and amyloid-β, such as Alzheimer's disease (AD) and frontotemporal degeneration.

[0205] This invention has discovered that the binding selectivity of the present invention relative to α-synuclein aggregates is higher than that relative to Tau protein and amyloid β aggregates; while on the other hand, compared with the binding selectivity relative to α-synuclein aggregates, 2-((1E,3E)-4-(6-(11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol, 2-((1E,3E)-4-(6-(11)C-methylamino)pyridin-3-yl)but- 1,3-dienyl)benzo[d]thiazol-6-ol, 1-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol and 1-(18)F-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol have higher binding selectivity relative to Tau protein aggregates.

[0206] The diagnostic kit of the present invention comprises both the former (α-synuclein aggregates) and the latter (Tau protein aggregates), which have high binding selectivity relative to α-synuclein aggregates. Therefore, by comparing the detection results of the former (the amount and / or distribution of detected light and radiation) with the detection results of the latter, it is possible to distinguish which substance is present in each imaged region. Specifically, it is possible to distinguish between α-synuclein aggregates and Tau protein aggregates, and to quantify their respective amounts. Therefore, it is possible to diagnose diseases related to α-synuclein aggregates and / or diseases related to Tau protein aggregates with high precision. For example, for each of the binders of the present invention, and 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-ol, 2-((1E,3E)-4-(6-((11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-ol, the ratio of their binding to α-synuclein aggregates and their binding to Tau protein aggregates can be predetermined, and the ratio of light intensity and radiation intensity in each region of the brain of the test organism after being given the former and after being given the latter can be measured. In this way, the relationship between the predetermined ratio and the measured intensity ratio can be used to distinguish whether α-synuclein aggregates or Tau protein aggregates are present in the region, and the amount of each can be quantified.

[0207] Furthermore, the diagnostic kit of the present invention can be a diagnostic kit combined with an imaging agent for amyloid β. The diagnostic kit combining the diagnostic kit of the present invention with an imaging agent for amyloid β can be used to distinguish which substance is present in each imaged region; specifically, it can distinguish α-synuclein aggregates, Tau protein aggregates, and amyloid β aggregates, and can quantify their respective amounts. Therefore, it can diagnose diseases related to α-synuclein aggregates, diseases related to Tau protein aggregates, and / or diseases related to amyloid β with high precision.

[0208] One embodiment of the present invention is a medicine for the treatment or prevention of diseases related to brain accumulation, and also relates to a medicine administered according to an administration plan based on data on the amount and / or distribution of brain accumulation obtained using the aforementioned diagnostic kit.

[0209] <Optical Imaging Methods>

[0210] The optical imaging method of the present invention includes the following steps: irradiating the brain of a test organism, which has been given the binding agent of the present invention, with light of a first wavelength from outside the brain, and then detecting light of a second wavelength emitted from the brain that is different from the first wavelength.

[0211] After administering an effective amount of the binder to a test organism, the binder reaching the brain binds to α-synuclein aggregates present within the brain. Optical imaging (visualization) of the α-synuclein aggregates can be achieved by irradiating the brain of the administered test organism with light of a first wavelength used to excite the binder from outside the brain, and detecting light of a second wavelength (e.g., fluorescence) emitted from the binder within the brain.

[0212] Mammals can be cited as test organisms. Examples of mammals include humans, rats, mice, rabbits, guinea pigs, hamsters, monkeys, dogs, minks, or miniature pigs.

[0213] There are no particular limitations on the method of administration; for example, it can be administered orally, intravenously, or intraperitoneally. Intravenous or intraperitoneal administration is preferred. Intravenous administration is the most preferred. The preferred dosage is 0.01 mg / kg to 5 mg / kg, 0.05 mg / kg to 3 mg / kg, or 0.1 mg / kg to 1 mg / kg, with 0.1 mg / kg to 1 mg / kg being the most preferred.

[0214] <Radiation Imaging Methods>

[0215] The radiographic imaging method of the present invention includes the following steps:

[0216] The detection of radiation emitted from the brain of a test organism to which the above-mentioned binder has been administered, wherein the above-mentioned binder comprises a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof, wherein one or more atoms of the compound of formula (I) are radioactive isotopes of that atom.

[0217] When an effective amount of the binding agent is administered to a test organism, the binding agent reaching the brain binds to α-synuclein aggregates present in the brain. By detecting the radiation emitted from the binding agent in the brain, radiographic imaging (visualization) of the α-synuclein aggregates can be performed.

[0218] Examples of test organisms include mammals. Examples of mammals include humans, rats, mice, rabbits, guinea pigs, hamsters, monkeys, dogs, minks, or miniature pigs. Preferably, the mammal is a human.

[0219] The administration method is not particularly limited, and non-oral administration methods such as oral, intravenous, or intraperitoneal administration are also acceptable. Intravenous or intraperitoneal administration is preferred. Intravenous administration is the most preferred. The dosage is preferably 5 ng / kg to 5 mg / kg, more preferably 5 ng to 20 μg / kg. The radiation energy administered is preferably 37 MBq to 7.4 GBq per individual, more preferably 370 GBq to 3,700 GBq.

[0220] <Screening methods for drugs to treat or prevent diseases associated with alpha synuclein aggregates in the brain>

[0221] The screening method (hereinafter also referred to as the screening method) for the treatment or prevention of diseases related to α-synuclein aggregates in the brain of the present invention comprises the following steps:

[0222] Candidate substances are screened based on the differences in the amount and / or distribution of light or radiation detected in the test organism before and after the administration of the candidate substance using the aforementioned optical imaging method or the aforementioned radiographic imaging method.

[0223] Diseases associated with alpha synuclein aggregates in the brain are the same as those described in the section on "Diagnostic Kits for Diseases Related to Substances Accumulated in the Brain" above.

[0224] In addition, the test organisms and administration methods are the same as those described in the "Optical Imaging Methods" and "Radiation Imaging Methods" sections above.

[0225] For example, if the amount (intensity) of light and radiation, such as fluorescence, from the binder is reduced after administration of the candidate substance compared to before administration, the candidate substance may be used as a therapeutic compound for the disease or symptom.

[0226] In addition, by comparing the amount and / or distribution of light or radiation from the tested organism with that of other normal mammals, if the results after administration of the candidate compound are closer to those of normal mammals than before administration, then the candidate compound may be used as a therapeutic compound for the disease or symptom.

[0227] <Methods for quantifying or determining the accumulation of α-synuclein aggregates in the brain>

[0228] The method of the present invention for quantifying or determining the accumulation of α-synuclein aggregates in the brain comprises the following steps: irradiating the brain of a test organism to which the aforementioned binding agent has been administered, with light of a first wavelength from outside the brain, wherein the aforementioned binding agent comprises a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof; then detecting light of a second wavelength emitted from the brain, different from the first wavelength; and...

[0229] Based on the detected amount and / or distribution of light, the accumulation of α-synuclein aggregates in the brain can be quantified or determined. This method uses optical imaging to quantify or determine the accumulation of α-synuclein aggregates in the brain.

[0230] The test organisms and administration methods are the same as those described in the <Optical Imaging Methods> section above.

[0231] By calculating the difference between the amount and / or distribution of light detected in the test organism and that in other normal mammals, it is possible to quantify the accumulation of α-synuclein aggregates in the brain and determine whether there is an accumulation of α-synuclein aggregates in the brain.

[0232] In other embodiments, the method of the present invention for quantifying or determining the accumulation of α-synuclein aggregates in the brain includes the following steps:

[0233] The detection of radiation emitted from the brain of a test organism to which the aforementioned binder has been administered, wherein the aforementioned binder comprises a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof, wherein one or more atoms of the compound of formula (I) are radioactive isotopes of that atom, and,

[0234] Based on the detected amount and / or distribution of radiation, the accumulation of α-synuclein aggregates in the brain can be quantified or determined. This method uses radiographic imaging to quantify or determine the accumulation of α-synuclein aggregates in the brain.

[0235] The test organisms and administration methods are the same as those described in the <Radiation Imaging Methods> section above.

[0236] By calculating the difference between the amount and / or distribution of light or radiation detected in the test organism and that in other normal mammals, it is possible to quantify the accumulation of α-synuclein aggregates in the brain and determine whether there is an accumulation of α-synuclein aggregates in the brain.

[0237] <Classification of substances accumulated in the brain and methods for determining their accumulation>

[0238] The method of the present invention for classifying and determining the accumulation of substances in the brain includes:

[0239] The first step involves irradiating the brain of a test organism to which the aforementioned binder has been administered with light of a first wavelength from outside the brain, and then detecting light of a second wavelength emitted from the brain that is different from the first wavelength. The aforementioned binder comprises a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof; and

[0240] Step 2 involves exposing the brain of a test organism, which has been exposed at a different time than in Step 1, to light of a third wavelength selected from at least one of 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-ol and 1-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-yloxy)propane-2-ol, from outside the brain. Then, light of a fourth wavelength, different from the third wavelength, emitted from the brain is detected.

[0241] Based on the light quantity and / or distribution data detected in step 1 and step 2, the classification and accumulation of substances in the brain are determined. This method uses optical imaging to determine the classification and accumulation of substances in the brain.

[0242] The test organisms and administration methods are the same as those described in the <Optical Imaging Methods> section above.

[0243] Since the test includes a first step of detecting light emitted from the brain of a test organism that has been given a binding agent with high binding selectivity to α-synuclein aggregates, and a second step of detecting light emitted from the brain of a test organism that has been given the aforementioned substance with high binding selectivity to Tau protein aggregates, by comparing the detection results of the first step (the amount and / or distribution of the detected light) with the detection results of the second step, it is possible to distinguish whether the substance accumulated in the brain is α-synuclein aggregates and / or Tau protein aggregates and to determine their accumulation.

[0244] In other embodiments, the method of the present invention for classifying and determining the accumulation of substances in the brain includes:

[0245] Step 1 involves detecting radiation emitted from the brain of a test organism to which the aforementioned binder has been administered, wherein the binder comprises a compound of formula (I), a pharmaceutically permissible salt thereof, or a solvate thereof, wherein one or more atoms of the compound of formula (I) are radioactive isotopes of that atom; and

[0246] Step 2 involves detecting radiation emitted from the brain of the test organism that was exposed at a different time than in Step 1 to at least one of 2-((1E,3E)-4-(6-((11)C-methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-ol and 1-(18)F-fluoro-3-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazolyl-6-yloxy)propane-2-ol, and...

[0247] Based on the radiation quantity and / or distribution data detected in step 1 and step 2, the classification and accumulation of substances in the brain are determined. This method uses radiographic imaging to determine the classification and accumulation of substances in the brain.

[0248] The test organisms and administration methods were the same as those described in the section on radiographic imaging methods above.

[0249] Since the test includes a first step of detecting radiation emitted from the brain of a test organism that has been given a binding agent with high binding selectivity to α-synuclein aggregates, and a second step of detecting radiation emitted from the brain of a test organism that has been given the aforementioned substance with high binding selectivity to Tau protein aggregates, by comparing the detection results of the first step (the amount and / or distribution of the detected radiation) with the detection results of the second step, it is possible to distinguish whether the substance accumulated in the brain is α-synuclein aggregates and / or Tau protein aggregates and to determine their accumulation.

[0250] <α-synuclein aggregation inhibitor>

[0251] The α-synuclein aggregation inhibitor of the present invention (hereinafter also referred to as aggregation inhibitor) contains the compound represented by formula (I) above, its pharmaceutically permissible salt, or a solvate thereof. The compound represented by formula (I), its pharmaceutically permissible salt, and its solvate contained in the α-synuclein aggregation inhibitor of the present invention are the same as the compound represented by formula (I), its pharmaceutically permissible salt, and its solvate contained in the above-mentioned <α-synuclein aggregate binding agent> section.

[0252] As described in the embodiments below, the α-synuclein aggregation inhibitor of the present invention can inhibit the formation of α-synuclein aggregates. Therefore, when the α-synuclein aggregation inhibitor of the present invention is administered to a mammal, the aggregation inhibitor reaching the brain of the organism can inhibit the aggregation of α-synuclein present in the brain of the organism, thereby preventing or treating diseases in mammals caused by α-synuclein aggregates, such as Parkinson's disease, Lewy body dementia (DLB), and multiple system atrophy (MSA).

[0253] As described above, the compound represented by formula (I), its pharmaceutically permissible salt, and its solvates exhibit high binding selectivity relative to α-synuclein aggregates.

[0254] The compound represented by formula (I), its pharmaceutically permissible salt, and its solvates have a high binding selectivity to α-synuclein aggregates. Therefore, it can be inferred that the compound represented by formula (I), its pharmaceutically permissible salt, and its solvates will bind to α-synuclein aggregates during α-synuclein aggregation, thereby inhibiting and preventing α-synuclein aggregation.

[0255] Mammals to which aggregation inhibitors are administered include, for example, humans, rats, mice, rabbits, guinea pigs, hamsters, monkeys, dogs, minks, or miniature pigs.

[0256] There are no particular limitations on the administration method of aggregation inhibitors, such as oral administration, intravenous administration, or intraperitoneal administration. They can also be administered directly to the local area where α-synuclein aggregates are present (brain). Intravenous or intraperitoneal administration is preferred, with intravenous administration being the most preferred. The preferred dosage is 0.01 mg / kg to 5 mg / kg, 0.05 mg / kg to 3 mg / kg, or 0.1 mg / kg to 1 mg / kg, with 0.1 mg / kg to 1 mg / kg being the most preferred.

[0257] Example

[0258] The present invention will be described in detail below through embodiments, but the present invention is not limited to these embodiments.

[0259] The following compounds were used.

[0260]

[0261]

[0262] (Preparation for BF-227)

[0263] BF-227 (i.e., 2-(2-[2-dimethylaminothiazolyl-5-yl]vinyl)-6-(2-[fluoro]ethoxy)benzoxazole) is manufactured by NARD Research Institute, catalog number NP039-0.

[0264] (Synthesis of PBB3)

[0265] PBB3 (i.e., (2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzo[d]thiazol-6-ol) was synthesized by the method described in Synthesis Example 3 of Patent Document 1.

[0266] (Synthesis of C05-01)

[0267] C05-01 (i.e., ((E)-2-(4-(2-fluoro-6-(methylamino)pyridin-3-yl)but-1-en-3-ynyl)benzo[d]thiazolyl-6-ol) was synthesized by the method described in Synthesis Example 23 of Patent Document 1.

[0268] (Synthesis of C05-02)

[0269] C05-02 (i.e., ((E)-5-(4-(6-(aminomethyl)benzo[d]thiazolyl-2-yl)but-3-en-1-ynyl)-N-methylpyridine-2-amine) was synthesized by the method described in Synthesis Example 11 of Patent Document 1.

[0270] (Synthesis of C05-03)

[0271] C05-03 (i.e., ((E)-2-(4-(6-(methylamino)pyridin-3-yl)but-1-en-3-ynyl)benzo[d]thiazol-6-ol) was synthesized by the method described in Synthesis Example 10 of Patent Document 1.

[0272] (Synthesis of C05-05)

[0273] C05-05 (i.e., ((E)-1-fluoro-3-(2-(4-(6-(methylamino)pyridin-3-yl)but-1-en-3-ynyl)benzo[d]thiazolyl-6-yloxy)propane-2-ol) was synthesized by the method described in Synthesis Example 21 of Patent Document 1.

[0274] (Synthesis of C05-08)

[0275] C05-08 (i.e., ((E)-2-(4-(4-(dimethylamino)phenyl)but-1-en-3-ynyl)benzo[d]thiazol-6-ol) was synthesized by the method described in Synthesis Example 6 of Patent Document 1.

[0276] (Synthesis of C05-09)

[0277] C05-09 (i.e., ((E)-2-(4-(4-(methylamino)phenyl)but-1-ene-3-ynyl)benzo[d]thiazol-6-ol) was synthesized by the method described in Example 7 of the Synthesis of Patent Document 1.

[0278] (Synthesis of C06-01)

[0279] C06-01 (i.e., 2-((1E,3E)-4-(2-fluoro-6-(methylamino)pyridin-3-yl)but-1,3-dienyl)benzofuran-5-ol) was synthesized by the method described in Synthesis Example 24 of Patent Document 1.

[0280] (Synthesis of C05-04)

[0281] C05-04(TM2) is synthesized using the following method.

[0282]

[0283]

[0284]

[0285] (Step 1)

[0286] Compound 1 (5.01 g, manufactured by Tokyo Chemical Industry) was reacted with tert-butyl nitrite (t-BuONO) and CuBr2 in acetonitrile (MeCN) to give compound 2 (5.94 g, 87.5% yield). The obtained compound 2 solidified in storage.

[0287] (Step 2)

[0288] Compound 2 (5.93 g) was reacted sequentially with n-butyllithium (n-BuLi) and N,N-dimethylformamide (DMF) in tetrahydrofuran (THF) to give compound 5 as a pale yellow solid (3.78 g, 80.5% yield).

[0289] (Step 3)

[0290] Compound 5 (3.78 g) was reacted with malonic acid and piperidine in pyridine to give compound 6 as a yellowish-brown solid (3.71 g, 80.6% yield).

[0291] (Step 4)

[0292] Compound 6 (4.00 g) was reacted with Dess-Martin Periodinane (DMP) and tetraethylammonium bromide (TEAB) in dichloromethane (DCM) to give compound 7 as a yellow solid (yield 1.85 g, 40.3%).

[0293] (Step 10)

[0294] Compound 14 (5.00 g, manufactured by Combi-Blocks) was reacted with N-bromosuccinimide (MBS) in acetonitrile (MeCN) to give compound 15 (6.80 g, 79.8%) as a pale pink solid.

[0295] (Step 11)

[0296] Compound 15 (6.80 g) was reacted with di-tert-butyl dicarbonate (DIBOC) and sodium hexamethyldisilazane (NaHMDS) in tetrahydrofuran (THF) to give compound 16, which was a yellow oil (2.42 g, 23.3% yield).

[0297] (Step 12)

[0298] Compound 16 (2.75 g) was reacted with methyl iodine (MeI) and sodium hydride (NaH) in N,N-dimethylformamide (DMF) to give compound 17 (2.82 g, 97.8% yield).

[0299] (Step 13)

[0300] Compound 17 (2.81 g) was reacted with trimethylsilylacetylene in a mixture of triethylamine (TEA) and tetrahydrofuran (THF) in the presence of bis(triphenylphosphine)palladium(II)dichloro(Pd(PPh3)2Cl2), copper iodide (CuI), and triphenylphosphine (PPh3) to give compound 18 (2.50 g, 84.2% yield). Compound 18 was cured in storage.

[0301] (Step 14)

[0302] Compound 18 (2.49 g) was reacted with tetra-n-butylammonium fluoride (TBAF) in tetrahydrofuran (THF) to give compound 19, which was a slightly brownish oil (yield 1.85 g, 95.7%).

[0303] (Step 17)

[0304] In a mixture of triethylamine (TEA) and tetrahydrofuran (THF), in the presence of bis(triphenylphosphine)palladium(II)dichloro(Pd(PPh3)2Cl2), copper iodide (CuI), and triphenylphosphine (PPh3), compound 7 (213.0 mg) was reacted with compound 19 (261.8 mg) to give compound 21 as a pale yellow solid (yield 256.3 mg, 74.0%).

[0305] (Step 18)

[0306] Compound 21 (245.3 mg) was reacted with trifluoroacetic acid (TFA) to give a yellow solid C05-04(TM2) (yield 174.2 mg, 92.0%). The overall yield was 15%. The obtained C05-04... 1 The H NMR and ESI MS results are shown below.

[0307] 1H NMR(DMSO-d6,400MHz)δ:2.79(3H,brs),3.84(3H,s),6.40(1H,m),6.93(1H,d, J=16.0Hz),7.12(1H,m),7.21(1H,d,J=16.0Hz),7.58-7.68(3H,m),7.87(1H,m)

[0308] ESI MS:m / z found 340.09(Calcd for C 18 H 14 FN3OS[M+H] + 340.09)

[0309] (Synthesis of C06-03)

[0310] C06-03(TM1) is synthesized using the following method.

[0311]

[0312]

[0313] (Step 5)

[0314] Compound 8 (24.95 g, manufactured by Tokyo Chemical Industry Co., Ltd.) was reacted with ethyl bromoacetate and K2CO3 in N,N-dimethylformamide (DMF) to give compound 9 as a white solid (26.07 g, 72.2% yield).

[0315] (Step 6)

[0316] Compound 9 (5.72 g) was reacted with N-methoxy-N-methylamine hydrochloride and lithium bis(trimethylsilyl)amide (LHMDS) in tetrahydrofuran (THF) to give compound 10 (5.68 g, 93.0% yield).

[0317] (Step 7)

[0318] Compound 10 (5.68 g) was reacted with lithium aluminum hydride (LAH) in diethyl ether (Et2O) to give compound 11 as a yellow solid (3.80 g, 89.3% yield).

[0319] (Step 8)

[0320] Compound 11 (3.69 g) was reacted with malonic acid and piperidine in pyridine to give compound 12 as a pale yellow solid (yield 4.41 g, 96.5%).

[0321] (Step 9)

[0322] Compound 12 (4.40 g) was reacted with Des Martin reagent (DMP) and tetraethylammonium bromide (TEAB) in dichloromethane (DCM) to give compound 13 as a pale yellow solid (yield 4.49 g, 87.9%).

[0323] (Step 15)

[0324] In a mixture of triethylamine (TEA) and tetrahydrofuran (THF), in the presence of bis(triphenylphosphine)palladium(II)dichloro(Pd(PPh3)2Cl2), copper iodide (CuI), and triphenylphosphine, compound 13 (596.6 mg) was reacted with compound 19 (765.2 mg) to give compound 20 as a pale yellow solid (375.5 mg, 37.7% yield).

[0325] (Step 16)

[0326] Compound 20 (375.5 mg) was reacted with trifluoroacetic acid (TFA) to give a yellow solid C06-03(TM1) (yield 149.5 mg, 52.2%). The overall yield was 10%. The obtained C06-03... 1 The H NMR and ESI MS results are shown below.

[0327] 1 H NMR(DMSO-d6,400MHz)δ:2.78(3H,brs),3.78(3H,s),6.38(1H,m),6.51(1H,d,J=16.0Hz),6 .91(1H,m),6.95(1H,s),6.98(1H,d,J=16.0Hz),7.14(1H,m),7.44-7.49(2H,m),7.59(1H,m)

[0328] ESI MS:m / z found 323.12(Calcd for C 19 H 15 FN2O2[M+H] + 323.12)

[0329] (〔 18 (Synthesis of F]C05-01)

[0330] The following method is used to synthesize [ 18 F〕C05-01.

[0331]

[0332] make〔 18F] Fluoride ions were dissolved in a 50% acetonitrile solution (0.4 mL) containing K.222 (Kryptofix 222) (7.5 mg) and potassium carbonate (2.77 mg). This solution was introduced into the reaction vessel, and the vessel was heated under a nitrogen stream to allow the solvent to dry and solidify. Then, anhydrous acetonitrile (0.1 mL) was added for azeotropic distillation, followed by thorough drying of the reaction vessel. The following reactions were carried out under light protection. A DMSO solution (300 μL) containing (E)-(5-(4-(6-(ethoxymethoxy)benzo[d]thiazolyl)but-3-en-1-yn-1-yl)-6-nitropyridin-2-yl)(methyl)carbamate tert-butyl ester (i.e., precursor C05-01) (1 mg) was added to the reaction vessel. The reaction mixture was heated at 110 °C for 10 minutes. After the reaction vessel cooled, 0.5 mL of 4N hydrochloric acid was added, and hydrolysis was carried out at 110 °C for 10 minutes. After the reaction vessel cooled, 2 mL of 1N sodium acetate was added and stirred, followed by the addition of 500 μL of HPLC solvent. The mixture was purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG 8010 mm × 250 mm, acetonitrile / 50 mM ammonium acetate = 6 / 4, 4 mL / min). The equivalent of […] 18 The fraction of F]C05-01 was recovered into a flask containing ethanol (300 μL), 25% ascorbic acid (100 μL), and Tween 80 (100 μL), and the solvent was removed by distillation under reduced pressure. The residue was dissolved in physiological saline (2.5 mL, pH 7.4) to obtain the [[]] solution for injection. 18 F〕C05-01.

[0333] (〔 11 (C] Synthesis of C05-04)

[0334] The following method is used to synthesize [ 11 C〕C05-04.

[0335]

[0336] The following synthesis was carried out under light-shielding conditions. Iodine […] was added at room temperature. 11C]methane was added to a solution of dimethyl sulfoxide (DMSO) (300 μL) containing (E)-6-fluoro-5-(4-(6-methoxybenzo[d]thiazolyl-2-yl)but-3-en-1-yn-1-yl)pyridine-2-amine (i.e., precursor C05-04-1) (1.5 mg). The reaction mixture was heated at 120 °C for 5 minutes. After the reaction vessel cooled, HPLC solvent (500 μL) was added. The mixture was purified by HPLC (HPLC conditions: Waters Atlantis prep T310 mm × 150 mm, acetonitrile / 50 mM ammonium acetate = 4 / 6, 5 mL / min). The equivalent of [ 11 The fraction of C][C05-04 was recovered into a flask containing ethanol (300 μL), 25% ascorbic acid (100 μL), and Tween 80 (75 μL), and the solvent was removed by distillation under reduced pressure. The residue was dissolved in physiological saline (3 mL, pH 7.4) to obtain the […] as an injection solution. 11 C]C05-04. In addition, the precursor C05-04-1 is synthesized by a method similar to that used for the synthesis of C05-04(TM2) described above.

[0337] (〔 18 (Synthesis of F]C05-04)

[0338] The following method is used to synthesize [ 18 F〕C05-04.

[0339]

[0340] make〔 18F] Fluoride ions were dissolved in 0.4 mL of a 50% acetonitrile solution containing Kryptofix 222 (7.5 mg) and potassium carbonate (2.77 mg). This solution was introduced into the reaction vessel, and the vessel was heated under a nitrogen stream to allow the solvent to dry and solidify. Then, anhydrous acetonitrile (0.1 mL) was added for azeotropic distillation, followed by thorough drying of the reaction vessel. The following reactions were carried out under light protection. A 300 μL solution of DMSO containing (E)-(5-(4-(6-methoxybenzo[d]thiazolyl)but-3-en-1-yn-1-yl)-6-nitropyridin-2-yl)(methyl)carbamate (i.e., precursor C05-04-2) (1.5 mg) was added to the reaction vessel. The reaction mixture was heated at 110 °C for 10 minutes. After the reaction vessel cooled, 0.3 mL of 4N hydrochloric acid was added, and hydrolysis was carried out at 110 °C for 10 minutes. After the reaction vessel cooled, 0.6 mL of 2N sodium acetate was added and stirred, followed by the addition of HPLC solvent (500 μL). The mixture was purified by HPLC (HPLC conditions: Waters Atlantis prep T3 10 mm × 150 mm, acetonitrile / 50 mM ammonium acetate = 4 / 6, 5 mL / min). The equivalent of […] 18 The fraction of F]C05-04 was recovered into a flask containing ethanol (300 μL), 25% ascorbic acid (100 μL), and Tween 80 (75 μL), and the solvent was removed by distillation under reduced pressure. The residue was dissolved in physiological saline (3 mL, pH 7.4) to obtain the [F]C05-04 fraction for injection. 18 F]C05-04. The precursor C05-04-2 is synthesized by a method similar to that used for the synthesis of C05-04(TM2) described above.

[0341] (〔 11 (C] Synthesis of C06-03)

[0342] The following method is used to synthesize [ 11 C〕C06-03.

[0343]

[0344] The following synthesis was carried out under light-shielding conditions. Iodine […] was added at room temperature. 11[C] Methane was added to a solution of dimethyl sulfoxide (DMSO) (300 μL) containing (E)-6-fluoro-5-(4-(5-methoxybenzofuran-2-yl)but-3-en-1-yn-1-yl)pyridin-2-amine (i.e., precursor C06-03-1) (1.5 mg). The reaction mixture was heated at 120 °C for 5 minutes. After the reaction vessel cooled, HPLC solvent (500 μL) was added. The mixture was purified by HPLC (HPLC conditions: Waters Atlantis prep T310 mm × 150 mm, acetonitrile / 50 mM ammonium acetate = 4 / 6, 5 mL / min). The equivalent of [ 11 The fraction of C][C06-03 was recovered into a flask containing ethanol (300 μL), 25% ascorbic acid (100 μL), and Tween 80 (75 μL), and the solvent was removed by distillation under reduced pressure. The residue was dissolved in physiological saline (3 mL, pH 7.4) to obtain the […] as an injection solution. 11 C]C06-03. In addition, the precursor C06-03-1 is synthesized by a method similar to that used for the synthesis of C06-03(TM1) described above.

[0345] (〔 18 (Synthesis of F]C06-03)

[0346] The following method is used to synthesize [ 18 F〕C06-03.

[0347]

[0348] make〔 18F] Fluoride ions were dissolved in 0.4 mL of a 50% acetonitrile solution containing Kryptofix 222 (7.5 mg) and potassium carbonate (2.77 mg). This solution was introduced into the reaction vessel, and the vessel was heated under a nitrogen stream to allow the solvent to dry and solidify. Then, anhydrous acetonitrile (0.1 mL) was added and azeotropically distilled, followed by thorough drying of the reaction vessel. The following reactions were carried out under light protection. A 300 μL solution of DMSO containing (E)-(5-(4-(5-methoxybenzofuran-2-yl)but-3-en-1-yn-1-yl)-6-nitropyridin-2-yl)(methyl)carbamate tert-butyl ester (precursor C06-03-2) (1.5 mg) was added to the reaction vessel. The reaction mixture was heated at 110 °C for 10 minutes. After the reaction vessel cooled, 0.3 mL of 4N hydrochloric acid was added, and hydrolysis was carried out at 110 °C for 10 minutes. After the reaction vessel cooled, 0.6 mL of 2N sodium acetate was added and stirred, followed by the addition of HPLC solvent (500 μL). The mixture was purified by HPLC (HPLC conditions: Waters Atlantis prep T3 10 mm × 150 mm, acetonitrile / 50 mM ammonium acetate = 4 / 6, 5 mL / min). The equivalent of […] 18 The fraction of F]C06-03 was recovered into a flask containing ethanol (300 μL), 25% ascorbic acid (100 μL), and Tween 80 (75 μL), and the solvent was removed by distillation under reduced pressure. The residue was dissolved in physiological saline (3 mL, pH 7.4) to obtain the [F]C06-03 fraction for injection. 18 F]C06-03. C06-03-2 is synthesized by a method similar to that used in the synthesis of C06-03(TM1) described above.

[0349] <Optical Imaging of the Human Brain>

[0350] (Brain Anatomy)

[0351] Local autopsies were performed on patients with Lewy body dementia (DLB) and Alzheimer's disease (AD) to obtain the post-mortem human brain.

[0352] Frozen DLB tissue was sectioned to a thickness of 20 μm using a cryostat (HM560, Carl Zeiss).

[0353] In addition, AD brain tissue was fixed in 10% neutral buffered formalin, embedded in paraffin blocks, and sectioned to a thickness of 6 μm.

[0354] (Intra-tube fluorescence microscopy measurement)

[0355] Freshly frozen sections of formalin-fixed amygdala tissue from DLB patients and dewaxed sections of formalin-fixed paraffin-embedded middle frontal gyrus tissue from AD patients were used. The brain sections and 30 μM of the above compounds were incubated in 50% ethanol at room temperature for 30 minutes. The sections were then washed twice: 5 minutes with 50% ethanol followed by 3 minutes with ultrapure water. After mounting with embedding medium (VECTASHIELD H-1000, Vector Laboratories), images of the lesion accumulation areas on the sections were obtained using a fluorescence microscope (DM4000, Leica). The fluorescence images are shown in Figure 1. Figure 1A and Figure 1B As shown in the image. The fluorescence intensity of the lesion area and the non-lesion-forming area (background) was quantified using analysis software (Image J). The results are as follows. Figure 2 As shown.

[0356] As shown in Figure 1 and Figure 2 As shown, compounds C05-01, C05-02, C05-03, C05-04, C05-05, C05-08, C05-09, and C06-02, represented by formula (I), were confirmed to bind to α-synuclein aggregates formed in the brains of DLB ​​patients with a binding strength of PBB3 or higher. Furthermore, it was confirmed that these compounds represented by formula (I) bind more strongly to α-synuclein aggregates than they bind to Tau and amyloid β aggregates formed in the brains of AD patients. Additionally, the same results were obtained from intratube fluorescence microscopy measurements of the brains of MSA patients as those of DLB ​​patients.

[0357] Furthermore, considering that the selectivity relative to α-synuclein aggregates is higher than that relative to aggregates formed in the brains of AD patients, the compounds shown in formula (I) above are superior to compound C06-01, which has a butadiene structure. Additionally, it has been confirmed that BF-227, previously reported as a PET probe for synuclein lesions, primarily binds to amyloid β aggregates in AD, and that BF-227's binding relative to α-synuclein aggregates is weaker than that of the compounds shown in formula (I) above.

[0358] <Optical Imaging of the Mouse Brain>

[0359] (Establishment of a mouse model after inoculation with α-synuclein filaments)

[0360] First, E. coli was used to express recombinant protein from mouse α-synuclein, which was then extracted and cultured in test tubes, thereby forming insoluble α-synuclein aggregates.

[0361] Then, the α-synuclein aggregates were inoculated into the striatum of mice. The α-synuclein aggregates spread to surrounding areas via neural networks, and α-synuclein lesions were observed in the neocortex of the brain several months later.

[0362] The following method was used to inoculate α-synuclein aggregates into the striatum of mice. First, the hair on the heads of 9-week-old male C57BL / 6 mice anesthetized with 1.5% (v / v) isoflurane was removed, and the scalp was disinfected with isodine. Then, cerolacaine was applied, and the scalp was incised to expose the skull. Next, a hole was drilled in the skull at a position 0.05 mm in the anterior fontanelle and 2 mm laterally. Using a glass pipette, 3 μl of α-synuclein fiber solution (4 mg / ml mouse α-synuclein fiber dissolved in physiological saline) was injected to a depth of 2 μm. The scalp was then repositioned and sutured.

[0363] (Intra-tube fluorescence microscopy measurement)

[0364] The brains of mice inoculated with α-synuclein fibers were excised, sectioned, and subjected to fluorescence staining analysis. Specifically, the mouse brain sections inoculated with α-synuclein fibers and 30 μM of the compound were incubated in 20% ethanol at room temperature for 30 minutes. The sections were then washed twice: 5 minutes with 20% ethanol followed by 3 minutes with ultrapure water. After mounting the sections with a mounting medium (VECTASHIELD H-1000), images of α-synuclein aggregates were obtained using a fluorescence microscope (DM4000). The same sections were washed with phosphate buffer and then autoclaved to activate antigenicity. Immunohistochemical staining was performed using anti-phosphorylated α-synuclein monoclonal antibodies (pS129, abcam, ab59264) (1:1000). After mounting the sections with VECTASHIELD H-1000, images of the same regions were obtained using a fluorescence microscope (DM4000). The results are as follows. Figure 3 As shown. Figure 3 In the middle, the right side shows the results of staining with antiphospho-α synuclein antibody, and the left side shows the results of C05-05 fluorescent staining.

[0365] according to Figure 3 The results show that the compound C05-05 shown in formula (I) above will bind to the lesions caused by phosphorylated α-synuclein.

[0366] (Two-photon laser scanning fluorescence microscopy examination in vivo)

[0367] Six weeks after injection of 1.5% (v / v) isoflurane into α-synuclein fibrillary solution, model mice were anesthetized, and cranial windows were established according to the Seylaz-Tomita method (Tomita et al. J Cereb Blood Flow Metab 25, 858-67, 2005). Two weeks after cranial window establishment, mice were anesthetized with 1.5% (v / v) isoflurane, and 50 μl of a DMSO saline solution containing 0.05% CO5-05 or PBB3 (DMSO:saline = 1:1) was administered via the tail vein. Ninety minutes after CO5-05 administration, or 30 minutes after PBB3 administration, the mice were fixed under a two-photon laser fluorescence microscope, and 100 μl of a saline solution containing 5 mM sulfonylrhodamine 101 was administered intraperitoneally. Two-photon fluorescence imaging was then performed at an excitation wavelength of 900 nm. The detection wavelengths for C05-05 and PBB3 were 500–550 nm, and the detection wavelengths for sulfonylrhodamine 101 were 573–648 nm. Results are as follows: Figure 4 As shown.

[0368] like Figure 4 The results showed that after intravenous injection of compound C05-05 (Formula (I)), the compound reached the brain and even neurons, binding to various α-synuclein lesions, as observed by two-photon fluorescence microscopy. Therefore, it can be said that even when the density and total number of lesions are low and difficult to detect with PET, lesions can be detected by biological fluorescence imaging. On the other hand, intravenous injection of PBB3 was observed, but no lesions were detected, confirming that compound C05-05 (Formula (I) had a higher lesion detection contrast than PBB3.)

[0369] <In vivo PET (Positron Emission Tomography) Imaging>

[0370] PET scans were performed using a micro PET Focus 220 animal scanner (Siemens Medical Solutions), which provides 95 slices at a thickness of 0.851 mm (center thickness), a 19.0 cm axial field of view (FOV), and a 7.6 cm in-section FOV. C57BL / 6 mice were anesthetized with 1.5% (v / v) isoflurane prior to the scan. [The text abruptly ends here, likely due to an incomplete sentence or missing information.] 18Immediately after injecting F]C05-01 (a positron-emitting radiolabeled compound C05-01) (24.3 ± 0.7 MBq), a 90-minute emission scan was performed in 3D list mode with an energy window of 350–750 keV. To avoid racemization of the compound, the injection and scanning were performed in a darkroom. All list mode data were compiled into 3D sinograms, which were then transformed into 2D sinograms using Fourier-rebinding (frames: 10 × 1, 6 × 5, 5 × 10 minutes). Summed images were obtained 30–60 minutes and 60–90 minutes post-injection using maximum a posteriori reconstruction. Furthermore, dynamic images were reconstructed using a 0.5-mm Hanning filter and filter-corrected backprojection. Referring to an MRI (Magnetic Resonance Imaging) template and using PMOD image analysis software (PMOD Technologies), volumes of interest (VOI) were defined for the striatum and midbrain. Results are as follows: Figure 5 and Figure 6 As shown.

[0371] The compound C05-01, as shown in formula (I), was labeled with a positron-emitting radionuclide and then intravenously injected into normal mice. PET scans were performed, and the results were as follows: Figure 5 and Figure 6 As shown, C05-01 was confirmed to have appropriate brain reach and clearance rate from the brain, and C05-01 exhibited good properties as a probe for imaging (visualizing) α-synuclein aggregates in the brain of organisms in PET. Furthermore, Figure 5 The illustration shows intravenous injection. 18 Images of the normal mouse brain in two coronal planes and one midsagittal plane, taken 1-30 minutes after F]C05-01, were overlaid with standard brain MRI images. Additionally, Figure 6 It corresponds to the striatum and midbrain. 18 F]C05-01 Time-radiation curve after intravenous injection.

[0372] <Optical Imaging of the Mouse Brain>

[0373] (In vitro imaging of biological organisms)

[0374] Six weeks after injection of the same α-synuclein fiber solution as described in the section above, mouse models were anesthetized with 1.5% (v / v) isoflurane and intraperitoneally administered C05-05 (1.66 mg / kg) or BF-227 (1.66 mg / kg).

[0375] Brain tissue was harvested and frozen 90 minutes after C05-05 administration and 120 minutes after BF-227 administration. 20 μm thick frozen sections were prepared using a cryostat (HM560). After mounting the sections with embedding medium (VECTASHIELD H-1000), images of α-synuclein aggregates on the sections were obtained using a fluorescence microscope (DM4000).

[0376] In addition, the same section was fixed by immersion overnight in phosphate buffer containing 4% paraformaldehyde. The fixed brain was washed with running water and then autoclaved to activate antigenization. Immunohistochemical staining was performed using anti-phosphorylated α-synuclein monoclonal antibodies (pS129, abcam, ab59264) (1:1000). After mounting the sections with embedding medium (VECTASHIELDH-1000), images of the same regions as described above were obtained using a fluorescence microscope (DM4000). Results are as follows... Figure 7 As shown. Figure 7 (a) is the image projected onto C05-05. Figure 7 (b) is an image projected onto the BF-227.

[0377] like Figure 7 The results showed that after intravenous injection of C05-05 into mice inoculated with α-synuclein fibers, in vitro imaging revealed that the compound traveled into the brain and even into neurons, binding to various α-synuclein lesions. On the other hand, intravenous injection of BF-227 was observed, but no lesions were detected, confirming that the lesion detection contrast of compound C05-05 (as shown in formula (I)) was higher than that of BF-227.

[0378] <In vivo PET (Positron Emission Tomography) Imaging>

[0379] PET scans were performed using a micro PET Focus 220 animal scanner (Siemens Medical Solutions), which provides 95 slices at a thickness of 0.851 mm (center thickness), a 19.0 cm axial field of view (FOV), and a 7.6 cm intra-sectional FOV. Prior to scanning, mice inoculated with α-synuclein fibers and mice injected with saline (control) were anesthetized with 1.5% (v / v) isoflurane. Intravenous injection…18 Immediately after injection of F]C05-05 (a positron-emitting radiolabeled compound C05-05) (30.8 ± 0.4 MBq), a 120-minute radiographic scan was performed in 3D list mode with an energy window of 350–750 keV. To avoid photoracemification of the compound, the injection and scanning were performed in a darkroom. All list mode data were compiled into 3D sinograms, which were then transformed into 2D sinograms using Fourier-rebinding (frames: 10 × 1, 6 × 5, 5 × 10 minutes). Summed images were obtained at 0–30 minutes, 30–60 minutes, 60–90 minutes, and 90–120 minutes post-injection using maximum a posteriori (MAP) reconstruction. Furthermore, dynamic images were reconstructed using a 0.5-mm Hanning filter and a filter-corrected backprojection method. Referring to the MRI template and using PMOD image analysis software (PMOD Technologies), the volume of interest (VOI) was set for the striatum and cerebellum.

[0380] The results are as follows Figures 8-12 As shown. Figure 8 (a) is [ 18 Results of mouse models 60–90 minutes after intravenous injection of F]C05-05 and injection of α-synuclein fibrillary solution. Figure 8 (b) shows the results in mice after saline injection. The top images show the superimposed images of the coronal plane of the brain including the striatum with standard brain MRI images, while the bottom images show the superimposed images of the coronal plane of the brain including the cerebellum with standard brain MRI images. Additionally, Figure 9 It corresponds to the striatum. 18 F]C05-05 intravenous injection time-radiimetry curve Figure 10 It corresponds to the cerebellum. 18 F]C05-05 intravenous injection time-radiimetry curve. Figure 11 It shows the striatum relative to [ 18 F〕The standardized uptake value ratio (SUVR) of the cerebellum over time after intravenous injection of C05-05. Figure 12The comparison is based on the average SUVR over the 90–120 minute timeframe between mice injected with α-synuclein fibrin solution and mice injected with saline (n = 4, **p < 0.005 by Student's t-test, mean ± SEMs).

[0381] The compound C05-05, as shown in formula (I), was labeled with a positron-emitting radionuclide and intravenously injected into mice modeling α-synuclein fibrils after injection of a solution and mice modeling α-synuclein fibrils after injection of saline (control). PET scans were performed, and the results showed appropriate brain accessibility and clearance rate from the brain. Furthermore, in the mice modeling α-synuclein fibrils after injection of the solution, […]. 18 F]C05-05 was clearly aggregated in the striatum of lesions rich in α-synuclein, confirming [ 18 F]C05-05 can be used to visualize α-synuclein aggregates in PET.

[0382] <The inhibitory effect of the compound shown in formula (I) on the aggregation of α-synuclein>

[0383] Recombinant protein, derived from A53T mutant human α-synuclein, was expressed in *E. coli* and extracted. The protein was then cultured in vitro to prepare α-synuclein aggregate seeds. 0.02 μg of the α-synuclein aggregate seeds were added to C05-05 (its DMSO solution) at a final concentration of 20 μM or 200 μM and stirred. A control experiment was conducted with only DMSO added. These samples were suspended in 50 μL of 1 mg / mL A53T mutant human α-synuclein monomer solution and incubated at 37°C with shaking for 1 day. 10 μL of the suspension was mixed with 90 μL of 1% sarcosine and ultracentrifuged at 100 K rpm for 20 min. The resulting precipitate was electrophoresed and then CBB stained onto the gel. Images of the gel were acquired using an imaging system, and the bands were quantified using analysis software (Image J). Results are as follows: Figure 13 and 14 As shown. Figure 13 These are the observation results of insoluble α-synuclein aggregates (indicated by arrows) contained in the insoluble fraction of sarcosine under various conditions. Figure 14 This is a bar chart with the control experiment (DMSO) set to 100%.

[0384] like Figure 14As shown, when a final concentration of 20 μM C05-05 was added to the α-synuclein aggregate seeds, the formation of insoluble α-synuclein aggregates decreased by approximately 20% compared to the control experiment. When a final concentration of 200 μM C05-05 was added, the formation of insoluble α-synuclein aggregates decreased by approximately 60% compared to the control experiment. Therefore, C05-05 has an inhibitory effect on α-synuclein aggregation. (ANOVA F(2,3)=51.57,p=0.0048,*p<0.05and**p<0.005by Bonferroni's post hoc test,mean±SEMs).

Claims

1. The use of the binder in compositions for optical imaging to prepare α-synuclein aggregates, wherein, The binder comprises a compound of formula (I) or a pharmaceutically permissible salt thereof. In formula (I), R1 and R2 are each independently selected from hydrogen and alkyl groups. R3 is hydrogen or halogen. Ring A is a benzene ring or a pyridine ring. Ring B is represented by either equation (i) or equation (ii). R4 and R5 are each independently selected from hydrogen, hydroxyl, alkoxy, haloalkoxy, halohydroxyalkoxy, and aminoalkyl.

2. The use according to claim 1, wherein, Ring A is a pyridine ring.

3. The use according to claim 1, wherein, Ring B is equation (i).

4. The use according to claim 1, wherein, The compounds represented by formula (I) are selected from the following group:

5. Use of the binder in compositions for radiographic imaging preparation of α-synuclein aggregates, wherein, The binder comprises a compound of formula (I) or a pharmaceutically permissible salt thereof. In formula (I), R1 and R2 are each independently selected from hydrogen and alkyl groups. R3 is hydrogen or halogen. Ring A is a benzene ring or a pyridine ring. Ring B is represented by either equation (i) or equation (ii). R4 and R5 are each independently selected from hydrogen, hydroxyl, alkoxy, haloalkoxy, halohydroxyalkoxy, and aminoalkyl; One or more atoms of the compound represented by formula (I) are radioactive isotopes of that atom.

6. The use according to claim 5, wherein, Ring A is a pyridine ring.

7. The use according to claim 5, wherein, Ring B is equation (i).

8. The use according to claim 5, wherein, The compounds represented by formula (I) are selected from the following group:

9. Use of the compound of formula (I) or a pharmaceutically permissible salt thereof in the preparation of a binder for optical imaging of α-synuclein aggregates in the brain. in, R1, R2, R3, R4, R5, ring A, and ring B in the compound represented by formula (I) are as defined in claim 1; Optical imaging involves irradiating the brain of a test organism to which the binder has been applied with light from outside the brain with light of a first wavelength, and then detecting light of a second wavelength, different from the first wavelength, emitted from the brain.

10. Use of the compound of formula (I) or a pharmaceutically permissible salt thereof in the preparation of a binder for radiographic imaging of α-synuclein aggregates in the brain. in, R1, R2, R3, R4, R5, ring A and ring B in the compound represented by formula (I) are as defined in claim 5, and one or more atoms of the compound represented by formula (I) are radioactive isotopes of that atom; Radiographic imaging includes detecting radiation emitted from the brain of a test organism to which the binder has been administered.

Images